鱼类NCCRP-1介导的NCC活性调控机理研究

非特异性细胞毒性细胞（NCC）是哺乳动物自然杀伤细胞的前体细胞，在鱼类先天性免疫中发挥着重要作用。非特异性细胞毒性细胞受体Ⅰ型基因（NCCRP-1）是该类细胞膜上关键的受体蛋白，是NCC发挥功能的分子基础，然而目前对于NCCRP-1介导的NCC活性调控网络与作用机理还缺乏系统的研究。本项目首先通过酵母双杂交、免疫共沉淀与pull-down等方法筛选与鉴定NCCRP-1的互作蛋白，随后采用显性抑制、western blot、real-time PCR等技术分析NCCRP-1与互作蛋白的作用机理及其对NCC胞内信号转导、效应分子表达及对细胞杀伤活性的影响，从而解析NCCRP-1对NCC活性调控的分子机理。研究结果不仅能丰富鱼类免疫学知识，也为深入了解脊椎动物细胞免疫的进化规律提供了新的方向。

Nonspecific cytotoxic cells (NCC) are precursor cells of mammalian natural killer cells, play important roles in fish innate immunity of. Nonspecific cytotoxic cell receptor type 1 gene (NCCRP-1) is a key receptor protein on NCC membrane, which is crucial for NCC activity. However, the regulatory network and molecular mechanism of NCC mediated by NCCRP-1 are poorly understood. In this project, interacting proteins of NCCRP-1 are screened and identified through yeast two hybrid, co-immunoprecipitation and pull-down method, etc. Then the interactions and mechanism of NCCRP-1 and target proteins are confirmed by dominant negative, western blot, real-time PCR analysis, etc. And the effects of the interactions on intracellular signal transduction, effector molecules expressions and NCC cytotoxicity are also investigated. These results will reveal the NCCRP1-modulated regulatory mechanisms on NCC activity. And the investigation will be not only to rich the knowledge of fish immunology, but also provide new ideas for the better understanding the evolution of vertebrate cellular immunity.